Balasiddaiah Anangi, Davanian Haleh, Aleman Soo, Pasetto Anna, Frelin Lars, Sällberg Matti, Lohmann Volker, Koh Sarene, Bertoletti Antonio, Chen Margaret
Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden.
Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.
J Virol. 2017 Apr 13;91(9). doi: 10.1128/JVI.00010-17. Print 2017 May 1.
Therapy with genetically modified autologous T cells has shown great promise in cancer therapy. For an efficient control of hepatitis C virus (HCV) infection, cytotoxic T cells (CTL) are pivotal, but persistence of activated T cells may lead to liver toxicity. Here, anti-HCV T cell receptors (TCRs) recognizing the HCV nonstructural (NS) NS3 or NS5 viral peptide target were examined by mRNA transfection of human peripheral blood lymphocytes (PBLs) derived from healthy donors as well as chronically infected HCV patients. Immunological analysis shows that while the CTLs expressing the NS5-specific TCR reduced HCV RNA replication by a noncytotoxic mechanism, the NS3-specific TCR-redirected CTLs were polyfunctional and inhibited HCV RNA replication through antigen-specific cytotoxicity. Transcriptome signatures from these two types of CTL responses revealed uniquely expressed gene clusters upon encountering hepatoma target cells presenting endogenously expressed HCV proteins. The NS3 TCR induced a rapid expression of apoptotic signaling pathways and formation of embryonic gene clusters, whereas the NS5A TCR activation induced extended proliferative and metabolic pathways as the HCV target cells survived. Our results provide detailed insights into basic HCV T cell immunology and have clinical relevance for redirecting T cells to target virally infected hepatoma cells. Due to the protective ability of HCV-specific T cells and the hepatotoxic potential that they possess, there is a great need for the understanding of the functional aspects of HCV-specific T cells. To circumvent the low level of precursor frequency in patients, we engineered primary CD8 T cells by mRNA TCR vectors to confer HCV specificity to new T cells. HCV TCRs that differ in antigen specificity and polyfunctionality were examined. mRNA TCR engineering of peripheral blood lymphocytes from healthy donors or chronically infected HCV patients resulted in strikingly high levels of HCV TCR expression and HCV-specific responses. While a cytotoxicity response from a polyfunctional T cell activation caused hepatotoxicity and the rapid induction of apoptotic signaling pathways, the noncytotoxic T cell activation showed extended proliferative, metabolic pathways and persistence of HCV target cells. Our results provide detailed insights into basic HCV T cell immunology and have clinical relevance for immune protection of HCV-associated diseases.
基因改造的自体T细胞疗法在癌症治疗中显示出巨大的潜力。对于有效控制丙型肝炎病毒（HCV）感染，细胞毒性T细胞（CTL）至关重要，但活化T细胞的持续存在可能导致肝毒性。在此，通过对来自健康供体以及慢性感染HCV患者的人外周血淋巴细胞（PBL）进行mRNA转染，检测了识别HCV非结构（NS）NS3或NS5病毒肽靶点的抗HCV T细胞受体（TCR）。免疫分析表明，表达NS5特异性TCR的CTL通过非细胞毒性机制降低了HCV RNA复制，而NS3特异性TCR重定向的CTL具有多种功能，并通过抗原特异性细胞毒性抑制HCV RNA复制。这两种CTL反应的转录组特征揭示了在遇到呈现内源性表达的HCV蛋白的肝癌靶细胞时独特表达的基因簇。NS3 TCR诱导凋亡信号通路的快速表达和胚胎基因簇的形成，而NS5A TCR激活诱导了扩展的增殖和代谢通路，因为HCV靶细胞存活了下来。我们的结果为HCV T细胞基础免疫学提供了详细的见解，并且对于将T细胞重定向以靶向病毒感染的肝癌细胞具有临床意义。由于HCV特异性T细胞的保护能力及其所具有的肝毒性潜力，非常需要了解HCV特异性T细胞的功能方面。为了规避患者中前体频率较低的问题，我们通过mRNA TCR载体对原代CD8 T细胞进行工程改造，赋予新T细胞HCV特异性。检测了抗原特异性和多功能性不同的HCV TCR。对健康供体或慢性感染HCV患者的外周血淋巴细胞进行mRNA TCR工程改造，导致HCV TCR表达水平和HCV特异性反应显著升高。虽然多功能T细胞激活引起的细胞毒性反应导致肝毒性并快速诱导凋亡信号通路，但非细胞毒性T细胞激活显示出扩展的增殖、代谢通路以及HCV靶细胞的持续存在。我们的结果为HCV T细胞基础免疫学提供了详细的见解，并且对于HCV相关疾病的免疫保护具有临床意义。
